A comparative study of the accumulation and detoxification of copper and zinc in Chlamydomonas reinhardtii: The role of extracellular polymeric substances

Extracellular polymeric substances (EPS) form an interface between microalgae and the surrounding water environment. Copper (Cu) and zinc (Zn) are essential micronutrients but may negatively affect microbial growth when their concentrations reach toxic thresholds. However, how EPS affect the accumulation and resistance of Cu and Zn in microalgae remains largely unknown. Here, we investigated EPS production upon Cu/Zn exposure and compared the tolerance strategies to the two metals by Chlamydomonas reinhardtii with and without EPS. Microalgal EPS synthesis was induced by Cu/Zn treatments, and the functional groups of polysaccharides and proteins were involved in complexation with metal ions. The extraction of EPS aggravated the toxicity and reduced the removal of metals from solution, but the effect was more pronounced for Cu than for Zn. Copper bound on the cell surface accounted for 54.6 ± 2.0 % of the Cu accumulated by C. reinhardtii, whose EPS components strongly correlated with Cu adsorption. In contrast, 74.3 ± 3.0 % of accumulated Zn was absorbed in cells, and glutathione synthesis was significantly induced. Redundancy and linear correlation analyses showed that the polysaccharide, protein and DNA contents in EPS were significantly correlated with Cu accumulation, absorption and adsorption but not with Zn. Data fitted to a Michaelis-Menten model further showed that the EPS-intact cells had higher binding capacity for Cu2+ but not for Zn2+. These differential impacts of EPS on Cu/Zn sorption and detoxification contribute to a more comprehensive understanding of the roles of microalgal EPS in the biogeochemical cycle of metals.